The MC1R gene is related to coat color in cattle and to lactoprotein content in milk. Polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing were used to analyze the MC1R of China Holstein Black-white Cattle, China Holstein Red-white Cattle, Luxi Yellow Cattle and Bohai Black Cattle. Three alleles (E^D, E⁺ and e) were identified. Most China Holstein Black-white Cattle had E^D and E⁺ alleles (E^D = 0.12, E⁺ = 0.80). The same was true for Bohai Black Cattle (E^D = 0.52, E⁺ = 0.47). On the other hand, most China Holstein Red-white Cattle and Luxi Yellow Cattle had the e allele (e/e = 0.95). E⁺/e genotype was present in China Holstein Red-white Cattle and Luxi Yellow Cattle. We reasoned that E^D and E⁺ lad to the synthesis of the black pigment. Furthermore, we found an important SNP (Single Nucleotide Polymorphism) at 725 position of the MC1R coding region.